High-throughput quantitation of SARS-CoV-2 antibodies in a single-dilution homogeneous assay

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1. Version published to 10.1038/s41598-021-91300-5

   Jun 10, 2021
2. 

   ScreenIT

   Mar 1, 2021

   SciScore for 10.1101/2020.09.16.20195446: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: All animal procedures were approved by the Institutional Animal Care and Use Committee of the Georgia Institute of Technology. IRB: Residual specimen materials were used for diagnostics development under a protocol that was reviewed and approved by the CDC Institutional Review Board§
   Randomization	not detected.
   Blinding	The serum samples from commercial sources were analyzed in a blinded fashion.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   The plates were washed 3×, and mouse secondary anti-human IgG antibody conjugated to HRP (Accurate Chemical …

   SciScore for 10.1101/2020.09.16.20195446: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: All animal procedures were approved by the Institutional Animal Care and Use Committee of the Georgia Institute of Technology. IRB: Residual specimen materials were used for diagnostics development under a protocol that was reviewed and approved by the CDC Institutional Review Board§
   Randomization	not detected.
   Blinding	The serum samples from commercial sources were analyzed in a blinded fashion.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   The plates were washed 3×, and mouse secondary anti-human IgG antibody conjugated to HRP (Accurate Chemical #JMH035098) was added at 1:10.000 dilution in 100 µL 5% milk in PBS-T and incubated for 30 min at RT.	anti-human IgG suggested: None
   Purification of IgG and IgM from human serum: Human serum samples from SARS-CoV-2 positive donors or control donors (N = 14 each) were pooled, and IgG and IgM antibodies were purified according to manufacturer’s recommendations using CaptureSelect CH1-Xl and Poros CaptureSelect IgM Affinity matrices (Thermo Fisher Scientific).	IgM suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   To express the proteins, human Expi293 cells (Thermo Scientific), growing in Expi293 Expression medium, were transfected with FectoPRO reagent (PolyPlus Transfection).	Expi293 suggested: RRID:CVCL_D615)
   Software and Algorithms
   Sentences	Resources
   Statistics: Statistical analyses were performed using GraphPad Prism software version 7.04 except for the kappa statistic, which was determined using the online version of GraphPad at https://www.graphpad.com/quickcalcs/kappa1.cfm (confidence interval calculated with equations by Fleiss).	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798) GraphPad suggested: (GraphPad Prism, RRID:SCR_002798)

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   Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

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   Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:

   One limitation of our study is that the work was conducted primarily to evaluate a new diagnostic approach. Control samples were sourced based on availability and not as part of a serological survey, and some associated data, including demographics, accurate location, symptoms, underlying conditions, and hospitalization history of the individuals were absent or incomplete and are not reported here. Furthermore, we note that the samples originated in the U.S., and would encourage testing representative control cohorts any time a serological test is implemented in a new area or population, or indeed species, to account for potential differences in baseline reactivity. Of note, we found that convalescent SARS-CoV-1 sera can also activate the assay (data not shown). While this has little practical relevance in human diagnostics due to the small number of convalescent SARS-CoV-1 patients, it is noteworthy with regards to ecological investigations and instances where infection with a closely related virus is a possibility. In summary, we used SARS-CoV-2 as an example to demonstrate that modern protein complementation enables design of simple yet robust serological assays that are easy to automate. Complementation detects antibodies due to their binding of multiple antigens simultaneously, and thus the assay has no apparent antibody class or species restrictions. The improved dynamic range means that more information can be derived from primary screening than is possible with ELISA....

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   Results from TrialIdentifier: No clinical trial numbers were referenced.

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   Results from Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

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   Results from JetFighter: We did not find any issues relating to colormaps.

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   Results from rtransparent:

   • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
   • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
   • No protocol registration statement was detected.

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3. 

   Version published to 10.1101/2020.09.16.20195446v1 on medRxiv

   Sep 18, 2020